Methods of genome installation in a recipient host cell

Patent metadata

Field	Value
Publication number	US-9481880-B2
Application number	US-59841408-A
Country	US
Kind code	B2
Filing date	May 1, 2008
Priority date	May 1, 2007
Publication date	Nov 1, 2016
Grant date	Nov 1, 2016

Abstract

The presently disclosed invention relates to methods of installing a genome isolated from one species (the donor) into suitably prepared cells of a second species (the recipient). Introduction of the donor genetic material into the recipient host cell effectively converts the recipient host cell into a new cell that, as a result of the operation of the donated genetic material, is functionally classified as belonging to the genus and species of the donor genetic material.

First claim

What is claimed is:

1. A method for transplantation of a Mycoplasma mycoides Large Colony donor cell genome in a recipient Mycoplasma capricolum , said method comprising:
(a) isolating a closed circular double stranded donor genome from a Mycoplasma mycoides Large Colony donor cell and suspending the isolated donor genome in agarose;
(b) deproteinizing the donor genome and liberating the donor genome from the agarose;
(c) preparing a Mycoplasma capricolum as a recipient cell for introduction of the donor genome by removing or inactivating the recipient cell genome, and simultaneously introducing the isolated donor genome from step (b) into the recipient cell in presence of polyethylene glycol, wherein the donor genome phenotypically transforms the recipient cell to that of the of Mycoplasma mycoides Large Colony.

2. The method of claim 1 , wherein the donor genome comprises a selection marker.

3. The method of claim 1 , wherein the donor genome encodes a restriction endonuclease that recognizes sequences in the recipient genome but not the donor genome.

4. The method of claim 1 , wherein the donor genome is methylated.

5. The method of claim 4 , wherein the donor genome is GATC Dam methylated.

Assignees

• Glass John I
• Alperovich Nina
• Hutchison Iii Clyde A
• Lartigue Carole
• Merryman Charles E
• Vashee Sanjay
• Venter J Craig
• Synthetic Genomics Inc

Inventors

• Glass John I
• Alperovich Nina
• Hutchison Iii Clyde A
• Lartigue Carole
• Merryman Charles E
• Vashee Sanjay
• Venter J Craig

Classifications

• C12N15/74

  Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora · CPC title

• C12N15/1079

  Screening libraries by altering the phenotype or phenotypic trait of the host (reporter assays C12N15/1086) · CPC title

• C12N15/10 Primary

  Processes for the isolation, preparation or purification of DNA or RNA (chemical preparation of DNA or RNA C07H21/00; preparation of non-structural polynucleotides from microorganisms or with enzymes C12P19/34) · CPC title